An ssDNA aptamer specific for detection and purification of hexahistidine-tagged proteins

doi:10.1016/j.ab.2020.113893

科研成果详情

题名	An ssDNA aptamer specific for detection and purification of hexahistidine-tagged proteins
作者	Wu, Shijia1; Liu, Yanting 2; Sun, Hongguang3; Zhong, Meizuo 4; Dai, Bichun3; Pan, Biyao 3; Shen, Zhijian5
发表日期	2020-10-15
发表期刊	ANALYTICAL BIOCHEMISTRY 影响因子和分区
语种	英语
原始文献类型	Article
关键词	Aptamer SELEX Hexahistidine-tag Protein purification
其他关键词	AFFINITY-CHROMATOGRAPHY ; LIQUID-CHROMATOGRAPHY ; RECOGNITION ; DNA ; MOLECULES ; LIGANDS
摘要	Aptamers are small-sized RNA or ssDNA ligands with a unique structure, which have high specificity and affinity to their cognate targets. Thus, in addition to the extensive values in various bio-medical fields, aptamers can also be alternatively used as affinity ligands in the bioprocess, such as for protein purification. In the present study, a hexahistidine specific aptamer named AptHis-C, was developed through the SELEX methodology, which has high affinity to hexahistidine, and its dissociation constant was as low as 20.8 nM. The structural prediction revealed that AptHis-C contains two connected stem-loop conformations. AptHis-C can only specifically recognize recombinant proteins with the hexahistidine-tag in simple or complex situations, and not to those with other tags. When immobilized on magnetic beads, AptHis-C can be used as a tool for hexahistidine-tagged recombinant protein purification. Its effectiveness is as good as traditional Ni-based beads. Besides, due to the intrinsic characteristics of nucleic acids, such as high thermal/chemical stability, immobilized aptamer-magnetic beads can be reused many times without an obvious decrease of purification effectiveness. This aptamer may represent a novel method for the detection and purification of hexahistidine-tagged recombinant proteins.
资助项目	First Affiliated Hospital of Wenzhou Medical University; Xiangya Hospital, Central South University
出版者	ACADEMIC PRESS INC ELSEVIER SCIENCE
出版地	SAN DIEGO
ISSN	0003-2697
EISSN	1096-0309
卷号	607 页码:113893
DOI	10.1016/j.ab.2020.113893
页数	8
WOS类目	Biochemical Research Methods ; Biochemistry & Molecular Biology ; Chemistry, Analytical
WOS研究方向	Biochemistry & Molecular Biology ; Chemistry
WOS记录号	WOS:000571726800005
收录类别	SCIE ; PUBMED ; EI ; SCOPUS
EI入藏号	20203409062704
EI主题词	Recombinant proteins
URL	查看原文
Pubmed记录号	32739349
Scopus记录号	2-s2.0-85089412842
通讯作者地址	[Shen, Zhijian]null
scopus学科分类	Biophysics;Biochemistry;Molecular Biology;Cell Biology
引用统计	被引频次：1[WOS] [WOS记录] [WOS相关记录]
文献类型	期刊论文
条目标识符	https://kms.wmu.edu.cn/handle/3ETUA0LF/7181
专题	第一临床医学院（信息与工程学院）、附属第一医院_浙江省胰脏肝脏危重性疾病重点实验室附属第一医院_血液内科
通讯作者	Shen, Zhijian
作者单位	1.Key Laboratory of Diagnosis and Treatment of Severe Hepato-Pancreatic Diseases of Zhejiang Province,The First Affiliated Hospital of Wenzhou Medical University,Wenzhou,China; 2.Department of Oncology,The First Affiliated Hospital of Xinxiang Medical University,Weihui,China; 3.Aptamer-Theranostics R&D Center,The First Affiliated Hospital of Wenzhou Medical University,Wenzhou,China; 4.Department of Oncology,Xiangya Hospital,Central South University,Changsha,China; 5.Department of Hematology,The First Affiliated Hospital of Wenzhou Medical University,Wenzhou,China
第一作者单位	附属第一医院; 第一临床医学院（信息与工程学院）、附属第一医院
第一作者的第一单位	附属第一医院
推荐引用方式 GB/T 7714	Wu, Shijia,Liu, Yanting,Sun, Hongguang,et al. An ssDNA aptamer specific for detection and purification of hexahistidine-tagged proteins[J]. ANALYTICAL BIOCHEMISTRY,2020,607:113893.
APA	Wu, Shijia., Liu, Yanting., Sun, Hongguang., Zhong, Meizuo., Dai, Bichun., ... & Shen, Zhijian. (2020). An ssDNA aptamer specific for detection and purification of hexahistidine-tagged proteins. ANALYTICAL BIOCHEMISTRY, 607, 113893.
MLA	Wu, Shijia,et al."An ssDNA aptamer specific for detection and purification of hexahistidine-tagged proteins".ANALYTICAL BIOCHEMISTRY 607(2020):113893.